Abstract
Background: In this investigation, we developed a new, rapid, novel HPLC technique for simultaneous routine analysis of amitriptyline and clomipramine. For this method, desipramine was used as an internal standard.
Experiment: Amitriptyline, clomipramine and desipramine (IS) were separated in a X-Terra RP-18 column (250mm×4.60mm ID×5µm) and DAD detector set at 210 nm. The composition of the mobile phase was acetonitrile:water 50:50 (v/v) at a flow rate of 1.0 mL min-1. Isocratic separation was performed in less than 6 min. Additionally, pKa values of studied compounds were evaluated by HPLC method in order to find the optimum pH value of separation. pKa values were determined at 45 and 50% (v/v) acetonitrilewater mixtures in order to find the effect of the mobile phase composition on the ionization constants. NLREG® program was used to calculate pKa values of drugs. The proposed method also has some advantages such as a number of cost and time saving benefits and can be routinely used for the analysis of pharmaceutical formulations. Result: The results show that developed method is suitable for the simultaneous analysis of these compounds in tablets without any interference from excipients.Keywords: Tricyclic antidepressants, amitriptyline, clomipramine, desipramine, pKa determination, liquid chromatography, method optimization.
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